Objectives:Clostridium difficile is a frequently identified cause of hospital-acquired infections. Identification of newly emerging hypervirulent C. difficile strains  like 027, 078 and 053 is mostly done by PCR-ribotyping. Although ribotyping is widely used the method lacks an interlaboratory interchangeable format. In order to overcome capillary-sequencer-dependent result variations of different types of sequencing machines a newly developed deterministic identification algorithm  AGES-WEBRIBO (webribo.ages.at)  was applied for capillary gel electrophoresis based Clostridium difficile-PCR ribotyping.

Methods: 2600 isolates collected between the years 2006 to 2009 at Austrian healthcare facilities were primarily analysed with an Applied Biosystems (ABI) 310 Genetic analyzer yielding 303 different PCR-ribotypes. 65 of the 303 ribotypes were randomly chosen and retested with on an ABI 3130 Genetic Analyzer. Quality control of data gained was done using the AGES-WEBRIBO internet platform.

Results: There was no increase in discrimatory power using different sequencer machines, although there was better narrow double-peak resolution with the ABI-3130 system. Three (4.5%) of the 65 on the ABI-3130 sequencer examined samples showed no correct identification after analysing the data with AGES-WEBRIBO.

Conclusion: 95.5% correct identifications of C. difficile ribotypes were achieved between different ABI-Sequencer models with the newly developed deterministic-data-analysis approach (AGES-WEBRIBO). This result clearly shows that every user is able to determine even rare PCR-ribotypes without a respective reference strain at hand. Further tests will show the ability of AGES-WEBRIBO (webribo.ages.at) to correctly identify data gained with capilliary sequencer models from other suppliers than ABI. We consider the updated version of AGES-WEBRIBO in combination with capillary gel electrophoresis based PCR ribotyping to be a universal tool for inter-laboratory comparability of C. difficile ribotyping.

Session Details

Date:

10/04/2010

Time:

00:00-00:00

Session name:

Abstracts 20th European Congress of Clinical Microbiology and Infectious Diseases